Identification of rat urinary glycoproteome captured by three lectins using gel and LC-based proteomics.
Many different types of urine proteome studies have been done, but urine glycoprotein studies are insufficient. Therefore, we studied the glycoproteins from rat urine, which could be used to identify biomarkers in an animal model. First, urinary proteins were prepared by using the dialysis and lyophilizing methods from rat urine. Glycoproteins enriched with lectin affinity purification, concanavalin A, jacalin and wheat germ agglutinin from the urinary proteins were separated by means of reverse-phase fast protein LC (FPLC) or 1-D PAGE. Each FPLC fraction and 1-D PAGE gel band were trypsin-digested and analyzed by means of nanoLC-MS/MS. LC-MS/MS analyses were carried out by using linear ion trap MS. A total of 318 rat urinary glycoproteins were identified from the FPLC fractions and gel bands; approximately 90% of identified proteins were confirmed as glycoproteins in Swiss-Prot. Many glycoproteins, known as biomarkers, including C-reactive protein, uromodulin, amyloid beta A4 protein, alpha-1-inhibitor 3, vitamin D-binding protein, kallikrein 3 and fetuin-A were identified in this study. By studying urinary glycoproteins collected from rat, these results may help to assist in identifying urinary biomarkers regarding various types of disease models.
Moon
Pyong-Gon
PG
Department of Molecular Medicine, School of Medicine, Kyungpook National University, Deagu, Republic of Korea.
Hwang
Hyun-Ho
HH
Boo
Yong Chool
YC
Kwon
Joseph
J
Cho
Je-Yoel
JY
Baek
Moon-Chang
MC
D000818:Animals; D015415:Biomarkers / Q000652:urine; D002853:Chromatography, Liquid; D003208:Concanavalin A; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000652:urine*; D008297:Male; D037121:Plant Lectins; D020543:Proteome / Q000032:analysis*; D051381:Rats; D053719:Tandem Mass Spectrometry; D014909:Wheat Germ Agglutinins
D015415:Biomarkers; D006023:Glycoproteins; D037121:Plant Lectins; D020543:Proteome; D014909:Wheat Germ Agglutinins; C043696:jacalin; D003208:Concanavalin A